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Master's Dissertation
DOI
10.11606/D.85.2004.tde-16042012-105910
Document
Author
Full name
Simone Aquino
E-mail
Institute/School/College
Knowledge Area
Date of Defense
Published
São Paulo, 2003
Supervisor
Committee
Villavicencio, Anna Lucia Casañas Haasis (President)
Arthur, Valter
Correa, Benedito
Title in Portuguese
Efeitos da radiação gama no crescimento de aspergillus flavus produtor de aflatoxinas e no emprego da técnica da reação em cadeia da polimerase (PCR) em amostras de grãos de milho inoculadas artificialmente
Keywords in Portuguese
aflatoxinas
Aspergillus
DNA
radiação gama
Abstract in Portuguese
O presente trabalho teve como objetivos verificar os efeitos da radiação gama em grãos de milho contaminados artificialmente com Aspergillus flavus Link produtor de aflatoxinas; demonstrar a aplicação da técnica da Reação em Cadeia da Polimerase (PCR) no diagnóstico de A. flavus, bem como verificar o efeito da radiação no perfil das bandas de DNA. Vinte amostras de grãos de milho com 200 g cada foram irradiadas individualmente com 20 kGy, para eliminar a contaminação microbiana. Em seguida, as amostras foram inoculadas com A. flavus toxigênico (1 x 106 esporos / ml), incubadas por 15 dias a 25 °C em ambiente com umidade relativa ao redor de 97,5% e irradiadas com 0; 2; 5 e 10 kGy. As amostras, 5 para cada dose de irradiação, foram analisadas individualmente quanto ao número de células fúngicas, atividade de água, teste de viabilidade (diacetato de fluoresceína e brometo de etídio), PCR e detecção de aflatoxinas (AFB). Os resultados demonstraram que as doses utilizadas foram efetivas na redução do número de Unidades Formadoras de Colônias (UFC/g), principalmente as doses de 5 e 10 kGy. Em adição, o teste de viabilidade mostrou uma diminuição de células viáveis com o aumento das doses de irradiação. A redução de AFB1 e AFB2 foi mais eficiente com o emprego de 2 kGy, comparativamente à dose de 5 kGy, enquanto a dose de 10 kGy degradou totalmente as aflatoxinas. Além disso, observou-se que AFB2 apresentou-se mais radiosensível. O emprego da técnica de PCR revelou a presença de bandas de DNA em todas as amostras.
Title in English
Effect of gamma radiation on the growth of Aspergillus flavus aflatoxins producer and on the use of polymerase chain reaction technique (PCR) in samples of maize grains artficially inoculated
Keywords in English
aflatoxins
Aspergillus
biological radiation effects
DNA
gamma radiation
maize
polymerase chain reaction
seeds
Abstract in English
The aim of this present study was to verify the effects of gamma radiation on the growth of Aspergillus flavus Link aflatoxins producer; to demonstrate the application of Polymerase Chain Reaction (PCR) technique in the diagnostic of A. Flavus, as well to verify the effect of radiation in the profile of DNA bands. Twenty samples of grains maize with 200 g each were individually irradiated with 20 kGy, to eliminate the microbial contamination. In following, the samples were inoculated with an toxigenic A. flavus (1x106 spores/ml), incubated for 15 days at 25 °C with a relative humidity of around 97,5% and irradiated with 0; 2; 5 and 10 kGy. The samples, 5 to each dose of irradiation, were individually analyzed for the number of fungal cells, water activity, viability test (fluorescein diacetate and ethidium bromide), PCR and aflatoxins (AFB) detection. The results showed that the doses used were effectives in reducing the number of Colony Forming Units (CFU/g) mainly the doses of 5 and 10 kGy. In addition, the viability test showed a decrease of viable cells with increase of irradiation doses. The reduction of AFB1 and AFB2, was more efficient with the use of 2 kGy in comparison with the dose of 5 kGy, while the dose of 10 kGy, degraded the aflatoxins. Thereby, it was observed that AFB2 showed to be more radiosensitive. The use of PCR technique showed the presence of DNA bands, in all samples
 
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2003AquinoEfeitos.pdf (1.67 Mbytes)
Publishing Date
2012-05-07
 
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